Deng Sheng-Guo, Deng Ze-Yuan, Fan Ya-Wei, Shan Bin, Xiong Dong-Mei
State Key Lab of Food Science and Technology, Institute for Advanced Study, Nanchang University, Nanchang 330047, China.
Guang Pu Xue Yu Guang Pu Fen Xi. 2010 Feb;30(2):476-80.
The interaction between astragalin (AST) from lotus leaf and deoxyribonucleic acid (DNA) in Tris-HCl buffer (pH = 7.4) was investigated by the application of fluorescence spectroscopy and ultraviolet absorption spectroscopy, the effects of ionic strength and anion quencher KI on the fluorescence intensity of AST from lotus leaf and the system of AST-DNA were explored, and the competitive binding to DNA between AST from lotus leaf and Neutral Red(NR)dye was also studied. The results demonstrated that AST could bind to DNA and the formed complex quenched the intrinsic fluorescence of AST from lotus leaf through static quenching mechanism. The quenching rate constants of biomolecule(Kq)of the reaction of DNA with AST from lotus leaf were calculated to be 3.120 X 10(12) and 2.630 X 10(12) L x mol(-1) x s(-1) by Stern-Volumer equation, the corresponding binding constants (Kd) were computed to be 3.412 x 10(12) and 1.762 x 10(4) L x mol(-1) and the number of binding sites(n) was counted to be 1.007 and 0.962 between AST from lotus leaf and DNA at 298 and 308 K, respectively. When bound to DNA, the AST from lotus leaf showed hypochromic effect and red shift in the absorption spectra. It was also found that different ionic strength had little or no effect on the fluorescence intensity of AST and AST-DNA, but the fluorescence intensity of AST-DNA quenched by anionic quencher KI was much less than that of free AST. AST could be intercalated into DNA and displaced the NR from the NR-DNA complex. It was showed that AST from lotus leaf could combine with DNA in the mode of intercalation.
采用荧光光谱法和紫外吸收光谱法研究了荷叶中紫云英苷(AST)与Tris-HCl缓冲液(pH = 7.4)中脱氧核糖核酸(DNA)的相互作用,探讨了离子强度和阴离子猝灭剂KI对荷叶AST荧光强度以及AST-DNA体系荧光强度的影响,还研究了荷叶AST与中性红(NR)染料对DNA的竞争结合作用。结果表明,AST能与DNA结合,形成的复合物通过静态猝灭机制猝灭荷叶AST的固有荧光。根据Stern-Volumer方程计算得出,荷叶AST与DNA反应的生物分子猝灭速率常数(Kq)分别为3.120×10¹²和2.630×10¹² L·mol⁻¹·s⁻¹,相应的结合常数(Kd)分别为3.412×10¹²和1.762×10⁴ L·mol⁻¹,在298 K和308 K时,荷叶AST与DNA之间的结合位点数量(n)分别为1.007和0.962。当与DNA结合时,荷叶AST在吸收光谱中表现出减色效应和红移。还发现不同离子强度对AST和AST-DNA的荧光强度影响很小或无影响,但阴离子猝灭剂KI猝灭的AST-DNA荧光强度远低于游离AST的荧光强度。AST可插入DNA中,并将NR从NR-DNA复合物中置换出来。结果表明,荷叶AST能以插入模式与DNA结合。